Department of Neurology, The First People's Hospital of Taizhou, No.218 Hengjie Road, Huangyan District, Taizhou, 318020, Zhejiang, China.
Neurochem Res. 2020 Dec;45(12):3034-3044. doi: 10.1007/s11064-020-03151-7. Epub 2020 Oct 23.
Temporal lobe epilepsy (TLE) is common intractable epilepsy that affects the patient's lives. The circular RNA circ_ANKMY2 (circ_ANKMY2) has been reported to be abnormally expressed in TLE. Nevertheless, the role and mechanism of circ_ANKMY2 in TLE are unclear. A human neuroblastoma cell line (SK-N-AS) was used for a series of studies. Expression levels of circ_ANKMY2, miR-106b-5p, and Forkhead Box Protein 1 (FOXP1) mRNA in TLE tissues were assessed through quantitative real-time polymerase chain reaction (qRT-PCR). Cell colony formation, proliferation, and apoptosis were determined by cell colony formation, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), or flow cytometry assays. The levels of FOXP1 protein, Ki67, B cell lymphoma (Bcl-2), Bcl-2 Associated X (Bax), and Cleaved caspase-3 were evaluated by western blot analysis. The relationship between circ_ANKMY2 or FOXP1 and miR-106b-5p was verified with dual-luciferase reporter assay. We observed that circ_ANKMY2 and FOXP1 expression were reduced while miR-106b-5p expression was increased in TLE tissues. Overexpression of circ_ANKMY2 decreased spontaneous recurrent seizures (SRSs) in rat TLE model and blocked cell colony formation, proliferation, and induced cell apoptosis in SK-N-AS cells. Importantly, circ_ANKMY2 was verified as a sponge for miR-106b-5p. In addition, miR-106b-5p mimics abolished circ_ANKMY2 elevation-mediated effects on colony formation, proliferation, and apoptosis of SK-N-AS cells. Also, FOXP1 served as a target for miR-106b-5p. And FOXP1 silencing overturned the effects of miR-106b-5p inhibitors on the colony formation, proliferation, and apoptosis of SK-N-AS cells. In sum, circ_ANKMY2 modulated TLE advancement via regulation of FOXP1 expression through sponging miR-106b-5p, and circ_ANKMY2 might be an underlying target for the improvement of TLE.
颞叶癫痫(TLE)是一种常见的难治性癫痫,影响患者的生活。已经报道环状 RNA circ_ANKMY2(circ_ANKMY2)在 TLE 中异常表达。然而,circ_ANKMY2 在 TLE 中的作用和机制尚不清楚。使用一系列研究的人神经母细胞瘤细胞系(SK-N-AS)。通过定量实时聚合酶链反应(qRT-PCR)评估 TLE 组织中 circ_ANKMY2、miR-106b-5p 和叉头框蛋白 1(FOXP1)mRNA 的表达水平。通过细胞集落形成、3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐(MTT)或流式细胞术测定细胞增殖和凋亡。通过蛋白质印迹分析评估 FOXP1 蛋白、Ki67、B 细胞淋巴瘤(Bcl-2)、Bcl-2 相关 X(Bax)和Cleaved caspase-3 的水平。通过双荧光素酶报告基因检测验证 circ_ANKMY2 或 FOXP1 与 miR-106b-5p 的关系。我们观察到 TLE 组织中 circ_ANKMY2 和 FOXP1 的表达降低,而 miR-106b-5p 的表达升高。circ_ANKMY2 的过表达降低了大鼠 TLE 模型中的自发性复发性癫痫(SRSs),并阻断了 SK-N-AS 细胞的集落形成、增殖,并诱导细胞凋亡。重要的是,circ_ANKMY2 被验证为 miR-106b-5p 的海绵体。此外,miR-106b-5p 模拟物消除了 circ_ANKMY2 升高介导的对 SK-N-AS 细胞集落形成、增殖和凋亡的影响。此外,FOXP1 是 miR-106b-5p 的靶标。FOXP1 沉默推翻了 miR-106b-5p 抑制剂对 SK-N-AS 细胞集落形成、增殖和凋亡的影响。总之,circ_ANKMY2 通过海绵 miR-106b-5p 调节 FOXP1 的表达来调节 TLE 的进展,circ_ANKMY2 可能是改善 TLE 的潜在靶点。